The long-term objective of this project is to provide a consistent supply of live, healthy squids for neuroscience research, primarily through the innovative approach of mariculture and secondarily by more effective methods of capturing and maintaining wild squids. Loliginid squids are renowned for their giant fiber system, which serves as a fundamental model for study of the nervous system and the cell membrane, and this project is intended to increase both the numerical and seasonal availability of live squids to the research community. The primary specific aim of this renewal is to culture the largest known loliginid squid, Loligo forbesi, from eggs to adult size and, through laboratory spawning, provide a source of eggs for the subsequent generation. This species has a distinguished history in biomedical research as well as superior qualities for culture, including very large hatching size, vigorous feeding, tolerance to stress, and behavioral adaptation to the laboratory environment. Our current culture experiment on Loligo forbesi and our recent achievement of culturing a smaller species, Loligo opalescens, through the life cycle indicates that providing Loligo forbesi through laboratory culture is an attainable goal. In order to accomplish this primary aim, experiments and evaluations will be undertaken on: acquisition and transport of squid eggs; fitness of hatchlings; food types and feeding densities for squids; rearing of food organisms; feeding, growth, behavior, survival, maturation and fecundity of cultured squids; and the effects of extrinsic factors on sexual maturation. Cultured squids will be fed a wide variety of live copepods, mysids, shrimps, and fishes. All squids and food organisms will be maintained or cultured in recirculating seawater systems (1,500 to 26,000 l capacity) in which water quality will be monitored closely; tank designs will be refined and enlarged, filtration capacities increased, and water management guidelines developed. Three local species will be captured and maintained on a seasonal basis. The small squid Lolliguncula brevis will be trawled year-round and supplied to researchers. Two larger species, Loligo plei and Loligo pealei, will be captured through night lighting on a seasonal basis to supplement the production of cultured Loligo forbesi. Throughout the grant period, the production schedule of Loligo forbesi and other laboratory cultured or wild-caught loliginid squids will be gradually increased so that squids of a wide size range will be available on a predictable basis to neuro-science investigators.